Assay Kit Name:
Creatine Kinase (CK) Activity Colorimetric Assay Kit
• Detection method- Absorbance (450 nm)
• Application- The kit can detect creatine kinase activity less than 1 mU.
• Serum & plasma.
• Animal tissues: muscle, brain, heart etc.
• Cell culture: Adherent or suspension cells.
Features & Benefits:
• Simple procedure; takes ~ 30-40 minutes
• Sensitive and high-throughput adaptable
• Kit contains all necessary reagents for measuring creatine kinase activity
• CK Assay Buffer
• CK Substrate
• ATP (Lyophilized)
• CK Enzyme Mix (Lyophilized)
• CK Developer (Lyophilized)
• NADH Standard (Lyophilized)
• Positive Control (Lyophilized)
Creatine Kinase (CK) also known as creatine phosphokinase (CPK) and ATP: creatine N-phosphotransferase is a common cellular enzyme (EC 220.127.116.11). It catalyzes the reversible conversion of creatine and ATP into ADP and phosphocreatine. CK is widely expressed in various tissues and cell types, with highest activity in striated muscles, heart tissue and brain. CK consists of two subunits: M (muscle) and B (brain), and has three isoenzymes: CK-MM (skeleton muscle), CK-MB (cardiac muscle), and CK-BB (brain). Increased CK level is associated with many diseases such as myocardial infarction, muscular dystrophy, pulmonary infarction and brain tumors. Accurate measurement of CK is crucial for early diagnosis, prediction and therapeutic strategy. In BioVision’s Creatine Kinase Activity Colorimetric Assay kit, creatine kinase converts creatine into phosphocreatine and ADP. The generated phoshocreatine and ADP reacts with CK Enzyme Mix to form an intermediate, which reduces a colorless Probe to a colored product with strong absorbance at 450 nm. The CK Activity Assay is high-throughput adaptable, simple and sensitive. This assay kit can detect Creatine Kinase activity less than 1 mU.
USAGE: For Research Use Only! Not For Use in Humans.
What is the sensitivity of this assay and how can plasma/whole blood samples be processed for this assay?
The assay has a detection sensitivity of 0.5 mU/ml of glutathione peroxidase in samples. The datasheet contains instructions for Erythrocytes. Whole blood can be processed similarly. Plasma can be diluted over a range and then the dilution that gives readings within the linear range of the standard curve can be used for the assay.
Are the assay buffers for K762 and K263 the same?
The assay buffers in K762 and K263 are different and we do not recommend swapping one with the other. The Assay buffer provides ideal pH, salt conditions for the enzymes in the assay and hence they are crucial for the success of the assay.
Which protein assay is compatible with this kit?
We suggest you use a detergent compatible BCA assay kit: http://www.biovision.com/bca-protein-assay-kit-ii-6783.html or http://www.biovision.com/extra-sense-bca-protein-assay-kit-6785.html
Is using fresh samples mandatory?
We always prefer fresh samples over frozen for the best results. However if needed, tissue can be flash-frozen in liquid nitrogen and then stored at -80C or cells/serum/plasma can be stored at -80C and used with this kit. We recommend minimizing freeze-thaw cycles for the best results.
What is the sample volume to be used with this kit for plasma samples from rat?
This depends on the amount of active CK enzyme in the sample. The sample volume per well would need to be optimized to make sure that the values obtained are within the linear range of the std. curve.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are the standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, the incubation time can be increased. As long as the standard curve is linear, it should be fine to use, since all the samples will also be measured under the same conditions on this curve.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.